Viral transformation for production of personalized type I interferons

Abstract

Type I interferons (IFN) are cytokines with many functions and have been widely used to treat many human diseases such as hepatitis C virus infection. Using the viral transformation and priming properties of Epstein-Barr virus, we have developed a system that can produce high levels of "personalized" IFNs, which are produced from the cells of the patient to whom the IFNs are to be administrated. We demonstrate the feasibility of the system. This seems to be the first report for the establishment of a personalized IFN-production system. The personalized IFNs could have a longer circulation time, fewer side effects but higher efficacy. We anticipate that the system can provide an improved form of IFN for medical uses.

Figure 1. Personalized type I IFN production system

Step 1: Blood collection. A small amount (5-10 ml) of blood will be drawn from an individual. The PBMC will be isolated from the blood. Step 2: EBV transformation. The PBMC will be infected by EBV, and the resulting immortalized B cells, lymphoblastoid cell lines (LCL), will grow to large volume in a tissue culture system. Step 3: IFN induction and purification. EBV-transformed cells are primed to produce robust levels of IFNs upon virus infection. IFNs can be purified. Step 4: IFN administration. IFNs can be administered back to the individual. Thus, the IFNs from a specific individual’s cells can be manufactured and consumed by the same person.

Figure 2. Generation of personalized IFNs in laboratory scale

A) The production of IFN-α by personalized cell lines. PBMCs were isolated from fresh buffy coats and EBV was inoculated with the PBMC (10⁷cells/ml) in the presence of cyclosporine in 96-well plates. The cells were fed by adding 50% of fresh medium at weekly intervals, and transformation was judged by outgrowth of cells over 3-5 weeks of culture. Personalized LCLs from three individuals (named Person A, B, or C) were infected by Sendai virus (Spafas, Inc; 200 HA units per ml) for seven hours. The IFN-α in supernatants was measured with the use of ELISA (PBL Biomedical Laboratories; catalog number 41100). Standard deviations are shown. +/−: with or without Sendai infection. B) Production of IFN-β from personalized cell lines. The personalized cell lines were infected by Sendai virus. Total RNAs were isolated and used for RPA with IFN-β and GAPDH probes. Specific protections of IFN-β and GAPDH RNAs are indicated. C) Comparable amounts of IFN-α can be produced from IB4 and Namalwa cell lines. IFNs are induced by infection of Sendai virus (200 HA units per ml) for seven hours. The cell culture media were collected, and the concentrations of IFN-α were measured with the use of ELISA. IFN-α production in IB4 was set as 100. IFN productions in PBMCs are varied among individuals.